不动杆菌对β-内酰胺类抗生素耐药机制及检测方法

近年来，随着临床广谱抗生素的大量使用，筛选出了各种各样的不动杆菌耐药株。不动杆菌对常用β-内酰胺类抗生素的耐药性也逐年增加。给治疗带来一定的困难。因此了解其耐药机理及其检测对防止耐药菌株的传播很有必要。     

鲍曼不动杆菌对喹诺酮类耐药机制

目的 探讨鲍曼不动杆菌对氟喹诺酮类药物的耐药机制。方法 采用K-B法检测35株鲍曼不动杆菌对氟喹诺酮类药物的药敏情况。十二烷基磺酸钠一聚丙酰胺凝胶电泳(SDS-PAGE)法检测鲍曼不动杆菌的外膜蛋白(OMP)，直接荧光法检测鲍曼不动杆菌对环丙沙星的吸收和积累，PCR法扩增gyrA基因并对其进行测序。结果 35株鲍曼不动杆菌中有20株对氟喹诺酮类药物耐药，耐药菌株与敏感菌株相比，外膜蛋白在约29Ku条带处消失，在26Ku条带处明显增强。耐药菌株药物积累量不及敏感菌株，经叠氮钠处理后，积累量上升并接近敏感菌株，对PCR扩增产物gyrA测序未发现有突变。结论 鲍曼不动杆菌对氟喹诺酮类药物的耐药与外膜蛋白的低通透性和主动外运有关。     

一种快速检测耐甲氧西林金黄色葡萄球菌方法的评价

目的评价胶体金法检测耐甲氧西林金黄色葡萄球菌(MRSA)和耐甲氧西林凝固酶阴性葡萄球菌(MRCNS)临床应用价值。方法采用头孢西丁纸片扩散法、PCR扩增mecA基因和胶体金法对临床分离的150株金黄色葡萄球菌(100株MRSA和50株MSSA)、68株溶血葡萄球菌(58株MRSH和10株MSSH)、135株表皮葡萄球菌(97株MRSE和38株MSSE)进行检测并作比较。结果 MRSA、MSSA、MRSE、MSSE、MSSH三种方法检测结果一致。58株MRSH中6株胶体金法检测阴性。结论胶体金法可作为MRSA简便而准确的检测方法,同时也可分析MRCNS。     

The clinical application of identification of Aspergillus spp.using ITS and β-tubulin gene sequences

Objective To study the clinical application of the ITS and β-tubulin gene regions in identification of Aspergillus spp. Methods One hundred and twenty-four Aspergillus strains that isolated from fungal rhino-sinusitis specimens were collected in Beijing Tongren Hospital, Capital Medical University from July 2007 to January 2010. They were identified by morphological and molecular methods. The first one included traditional culture, slide culture, and microscopic examination after lactophenol cotton blue stain and KOH digestion. The second one was amplifying and sequencing the part of ITS and β-tubulin gene and aligned all the sequences in the GenBank, European Molecular Biology Laboratory nucleotide sequence database, and DNA Data Bank of Japan. Results Of the 56 Aspergillus flavus identified by morphological features, fifty-five isolates were identified as Aspergillus flavus and 1 isolates was Aspergillus parasiticus by the ITS and β-tubulin gene region sequence analysis. In the 37 Aspergillus fumigatus identified by morphological method, and all the 37 isolates were identified as species complex of Aspergillus fumigatus by the ITS region sequence analysis, but through the sequence analysis of β-tubulin gene region, thirty-five isolates were identified as Aspergillus. fumigatus and 2 were Aspergillus lentulus. Twenty-one isolates were identified as Aspergillus versicolor by morphological method, but 16 of them were identified as Aspergillus. versicolor and 5 can not be identified to species level by the ITS region sequence. And by comparative-sequence analysis of β-tubulin gene region, the 5 isolates were identified as Aspergillus sydowii,the other 16 isolates were Aspergillus. versilcolor. Ten isolates were identified as Aspergillus nidulans by morphological features, the ITS and β-tubulin gene region sequence analysis. Conclusions β-tubulin gene sequencing is more suitable for identifying Aspergillus, and could identify Aspergillus spp. to species level Sequences of ITS region could only identify Aspergillus spp. to species complex.     

我院门诊处方点评及不合理用药分析

目的分析门诊处方不合理用药情况,促进临床合理用药。方法抽取2011年4-10月处方共1500张,按处方点评指标进行分析。结果 1 500张处方中,合格1 405张（93.7%）,不合格95张（6.3%）。抗菌药物占40.5%,注射剂占17.6%。结论处方点评使医院处方管理、处方开具和处方调剂更加科学、合理、规范。     

临床药师参与55例抗菌药物应用会诊情况分析

目的了解临床药师参与感染性疾病会诊的意见及其落实情况。方法对2011年5-11月笔者参与会诊的55例感染性疾病的会诊记录及会诊邀请单进行统计分析。结果 55例临床邀请临床药师会诊病例均有病程记录,其中49例明确提出更换抗菌药物,3例建议按临床医生原治疗方案继续治疗,3例提出注意事项及用药建议,采纳率为96%。结论临床药师应不断学习临床和药学知识,提高会诊质量,在疑难感染疾病治疗中发挥作用。     

注射用头孢哌酮钠舒巴坦钠致过敏性休克1例

患者,男,41岁,诊断为尿路感染,于2011年9月23日入院治疗。否认家族药物过敏史,既往药物过敏史不祥。给予头孢哌酮钠舒巴坦钠3.0g(立健舒,深圳立健制药有限公司生产)+生理盐水50ml(皮试阴性)静脉滴注。于26日凌晨2点许,患者静脉滴注头孢哌酮钠舒巴坦钠约10min时,突然出现胸闷、胸痛、气促,继而出现面色潮红,神情烦躁,大声叫喊,全身未见皮疹,无瘙痒。考虑为过敏性休克。立即停止输液,BP     

慢性鼻窦炎患者与健康人群鼻腔菌群分析

目的调查慢性鼻窦炎患者鼻道主要细菌的种类。方法选取2018年1月至12月收治入院的慢性鼻窦炎患者137例,健康成人111例。在鼻内镜引导下用鼻拭子采集鼻分泌物标本,行普通细菌培养、厌氧细菌培养和基质辅助激光解吸电离飞行时间质谱分析。结果健康成人组中需氧菌以凝固酶阴性葡萄球菌(44.7%)和棒状杆菌(38.5%)为主,少数人群有金黄色葡萄球菌(7.7%)、铜绿假单胞菌(2.3%)和革兰阴性杆菌(1.9%);单一细菌定植比例为12.6%,2种细菌定植比例为73.9%,3种细菌定植比例13.5%。患者中需氧细菌以金黄色葡萄球菌(24.0%)、流感嗜血杆菌(6.2%)、克雷伯菌属(6.8%)为主的肠杆菌科细菌和铜绿假单胞菌(4.3%)为主;厌氧菌的检出率明显高于健康人群;单一细菌感染占36.5%,2种细菌感染占48.9%,3种细菌感染占14.6%。金黄色葡萄球菌、肺炎克雷伯菌、流感嗜血杆菌、铜绿假单胞菌单一或合并厌氧菌感染时临床症状较重,容易形成细菌耐药,治疗周期长。两组间致病菌金黄色葡萄球菌、流感嗜血杆菌、克雷伯菌为主的肠杆菌科细菌的检出率差异有统计学意义(P0.05)。结论慢性鼻窦炎患者和健康成人鼻道的细菌类型有明显区别,临床应根据培养结果规范使用治疗药物。     

两种不同方法检测血清降钙素原的相关性分析

目的 采用快速上转发光法与免疫荧光法检测血清中的降钙素原,比较两种不同方法学的相关性.方法 随机选取100例常规送检血清样本,同时用降钙素原上转发光试剂和免疫荧光试剂进行检测,比较两组结果的一致性.结果 上转发光法降钙素原检测试剂与免疫荧光法降钙素原检测试剂的相关性为0.997,符合率为98％,相关性良好.结论 POCT上转发光快速试剂与荧光免疫试剂达到高度一致的检测结果,可以广泛应用于临床、急诊、移动救护领域等.     

临床分离副溶血弧菌药敏表型和毒力特征分析

目的 了解临床分离的副溶血弧菌的耐药性及毒力特征.方法 通过医生工作站获取患者临床资料并通过实验室信息管理系统(LIS)统计2009至2012年北京同仁医院肠道门诊细菌性腹泻病原菌种类及数量;纸片扩散法检测副溶血弧菌对15种抗生素的耐药表型;PCR技术检测直接溶血毒素(TDH)基因(tdh)、TDH-相关溶血毒素基因(trh)和两个Ⅲ型分泌系统2基因(T3SS2α、T3SS2β).结果 副溶血弧菌在细菌性腹泻中所占比例为37.90％(274/723),居于首位.216株副溶血弧菌对氨苄西林、头孢噻吩、头孢呋辛、阿米卡星4种抗生素的耐药率分别为100％ (216/216)、44.91％ (97/216)、24.07％(52/216)、9.26％(20/216).对另外11种抗生素敏感或中度敏感.216株副溶血弧菌中tdh携带率为96.76％(209/216),trh携带率为1.39％ (3/216);T3SS2携带率为100％,其中213株只携带T3SS2α,2株只携带T3SS2β,1株同时携带T3SS2α和T3SS2β.结论 副溶血弧菌对氨苄西林耐药率最高,对其他抗生素不同程度耐药.造成人类腹泻的副溶血弧菌主要是tdh阳性菌株;Ⅲ型分泌系统在菌株致病中起重要作用.